We will continue to characterize RNA polymerase II and compare this with polymerases I and II. Product analysis of III will be determined and the level of enzyme during the cell cycle determined. Employing nuclei and nucleohistones isolated at different stages of the cell cycle and effort will be continued to measure random or non-randomness of RNA transcription during the cell cycle. In the reconstructed system RNA polymerase I, II and III will all be used with isolated templates. Using specific antibodies and pulse labelling of cells, we will attempt to directly prove the time of de novo protein synthesis for a specific enzymes during the cell cycle. Further biochemical characterization of cell cycle, sporulation and outgrowth will be continued to provide the background technology and information to test preferential transcription of the genome during these developmental phases.